LOCUS       sequence-12156-e        6066 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6066)
  AUTHORS   Karginov AV, Ding F, Kota P, Dokholyan NV, Hahn KM
  TITLE     Engineered allosteric activation of kinases in living cells.
  JOURNAL   Nat Biotechnol. 2010 Jun 27. ():.
  PUBMED    20581846
REFERENCE   2  (bases 1 to 6066)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Biotechnol. 2010 Jun 27. ():."
COMMENT     Sequence Label: pCMV5-Flag-RapR-p38
FEATURES             Location/Qualifiers
     source          1..6066
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     127..149
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     141..158
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     142..158
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        319..698
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        699..902
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     853..873
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             970..993
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    2396..3018
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2595..2614)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3040..3060)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3045..3375
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3226..3361
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3413..3432)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3414..3432)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3447..3463)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3447..3463)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3460..3482)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3471..3487
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3495..3501,3502..3519,3520..3525))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3540..3561
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3678..3695)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3848..4436)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3928..3947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4607..5398,5399..5467))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5230..5249
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5468..5572)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5599..6027
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5686..5705)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5896..5917
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 cccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc
       61 tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag
      121 ggttttccca gtcacgacgt tgtaaaacga cggccagtgc caagctgatc tatacattga
      181 atcaatattg gcaattagcc atattagtca ttggttatat agcataaatc aatattggct
      241 attggccatt gcatacgttg tatctatatc ataatatgta catttatatt ggctcatgtc
      301 caatatgacc gccatgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
      361 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
      421 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
      481 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
      541 cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg
      601 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt
      661 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
      721 ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
      781 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc
      841 ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
      901 ctcgtttagt gaaccgtcag aattcagatc tggtaccacg cgtatcgata agcttcggca
      961 gccaacatgg actataagga cgatgatgac aaaatgtcgc aggagaggcc cacgttctac
     1021 cggcaggagc tgaacaagac catctgggag gtgcccgaac gataccagaa cctgtccccg
     1081 gtgggctcgg gcgcctatgg ctcggtgtgt gctgcttttg atacatacct gagcccaggc
     1141 acctgcgtgg tgcactacac cgggatgctt gaagatggaa agaaatttga ttcctcccgg
     1201 gacagaaaca agccctttaa gtttatgcta ggcaagcagg aggtgatccg aggctgggaa
     1261 gaaggggttg cccagatgag tgtgggtcag agagccaaac tgactatatc tccagattat
     1321 gcctatggtg ccactgggca cccaggcatc atcccaccac atgccactct cgtcttcgat
     1381 gtggagcttc taaaactgga aggcaatcca gctttgacgg ggcatcgtgt ggcagttaag
     1441 aagctgtcga gaccgtttca gtccatcatt cacgccaaaa ggacctaccg agagttgcgt
     1501 ctgctgaagc acatgaaaca cgaaaatgtg attggtctgt tggatgtgtt cacacccgca
     1561 aggtcactgg aggaattcaa tgacgtgtac ctggtgaccc atctcatggg ggcggacctg
     1621 aacaacatcg tgaagtgcca gaagctgacc gacgaccacg ttcagtttct catctaccag
     1681 atcctccgag ggctgaagta tatacattcg gctgacataa ttcacaggga cctaaagccc
     1741 agcaacctag ctgtgaacga agactgtgag ctcaagattc tggattttgg gctggctcgg
     1801 cacactgatg atgagatgac aggctacgtg gctaccaggt ggtaccgagc cccagagatc
     1861 atgctgaatt ggatgcacta taaccagaca gtggatattt ggtccgtggg ctgcatcatg
     1921 gctgagctgt tgaccggaag aacgttgttt cctggtacag accatattga tcagttgaag
     1981 ctcattttaa gactcgttgg aaccccaggg gctgagcttc tgaagaaaat ctcctcagag
     2041 tctgcaagaa actacattca gtctctggcc cagatgccga agatgaactt cgcaaatgta
     2101 tttattggtg ccaatcccct ggctgtcgac ctactggaga agatgctcgt tttggactca
     2161 gataagagga tcacagcagc ccaagctctt gcgcatgcct actttgctca gtaccacgac
     2221 cctgatgatg agcctgttgc tgacccttat gaccagtcct ttgaaagcag ggaccttctc
     2281 atagatgagt ggaagagcct gacctatgat gaagtcatca gctttgtgcc accacccctt
     2341 gaccaagaag aaatggagtc ctgagcatgc ctgcaggtcg actctagagg atcccgggtg
     2401 gcatccctgt gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc
     2461 caccagcctt gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta
     2521 taatattatg gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg
     2581 tagggcctgc ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac
     2641 tgcaatctcc gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg
     2701 attccaggca tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt
     2761 caccatattg gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc
     2821 tcccaaattg ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt
     2881 aaaataacta taccagcagg aggacgtcca gacacagcat aggctacctg ccatggccca
     2941 accggtggga catttgagtt gcttgcttgg cactgtcctc tcatgcgttg ggtccactca
     3001 gtagatgcct gttgaattgg gtacgcggcc agcttctgtg gaatgtgtgt cagttagggt
     3061 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3121 cagcaaccag gtgtggaaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3181 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3241 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3301 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3361 ctaggctttt gcaaaaagct cctcgaggaa ctgaaaaacc agaaagttaa ttccctatag
     3421 tgagtcgtat taaattcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     3481 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     3541 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     3601 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     3661 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3721 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3781 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaag gccgcgttgc
     3841 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     3901 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     3961 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4021 cgggaagcgt ggcgctttct caatgctcac gctgtaggta tctcagttcg gtgtaggtcg
     4081 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4141 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4201 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4261 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     4321 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4381 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4441 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4501 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4561 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4621 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4681 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4741 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     4801 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     4861 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     4921 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     4981 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5041 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5101 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5161 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5221 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5281 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5341 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5401 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5461 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5521 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5581 cccgaaaagt gccacctgac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     5641 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     5701 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cggggcatcc
     5761 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     5821 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     5881 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     5941 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6001 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacaaaatat taacgtttac
     6061 aatttc
//
